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We narrowed to 958 results for: ache

Showing: 421 - 440 of 958 results
  1. Pairing CombiGEM and CRISPR for Combinatorial Genetic Screening

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    Blog Post
    ...containing combinations of different gRNAs in each plasmid. Each gRNA combination can be tracked and quantitatively...continual advancements in gRNA design necessary to achieve maximal on-target and minimal off-target activities...lentiviral plasmids containing one or more gRNAs. As each ligation reaction uses a pool of gRNAs as starting...
  2. The Future of Research Symposium Boston 2015

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    Blog Post
    ...example, in 2010, less than 15% of postdocs had achieved tenure-track positions within 5-7 years of earning...BEST) awards, which are designed to support new approaches that address the need to train our young scientists...University School of Medicine and the University of Massachusetts Medical School. Graduate education programs ...PMCID: PMC4304227. 3. Sauermann, Henry, and Michael Roach. "Science PhD career preferences: levels, changes...
  3. New Tool for Lineage Tracing: The ClonTracer Library

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    Blog Post
    ...(MOI) such that each individual cell receives only one barcode. The abundance of each barcoded clone can...using NGS because the error rate of current NGS approaches allows for a detection limit of roughly 0.1%....populations can be eradicated up-front quickly. This approach is in contrast to the current treatment paradigm...
  4. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

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    Blog Post
    ...Christopher Voigt explains it, his lab at the Massachusetts Institute of Technology has been “working on...enough regulatory proteins that don’t interfere with each other. CRISPR is so orthogonal and programmable ...guide RNAs connected because the signal degrades at each layer. If we could fix these things, then I think...make orthogonal variation. The first one worked in each case. Sometimes in CRISPR people talk about off-...
  5. CRISPR 101: Cytosine and Adenine Base Editors

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    Blog Post
    ...target sites for base editing (Kim et al., 2017). For each base editor, they observed editing activity with...sites previously difficult to target. ABE8s can achieve 98-99% target modification in primary T cells, ...Grünewald et al., 2020). Another lab took a similar approach. Dali Li’s lab’s A&C-BEmax consists of a fusion...activity compared to ABEmax. In an alternative approach, engineering and directed evolution of the TadA...10.1038/s41587-022-01533-6 Nishida, K., Arazoe, T., Yachie, N., Banno, S., Kakimoto, M., Tabata, M., Mochizuki...
  6. Hot Plasmids - October 2020

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    Blog Post
    ...brain structure and function, with the advent of each tool expanding our ability to study and understand...traditionally introduced in Caenorhabditis elegans on extrachromosomal arrays can lead to artifacts due to overexpression... FP genes into C. elegans? By using a split FP approach. Split-wrmScarlet is a new split red FP that is... elegans cell lines to express wrmScarlet1-10 unattached to any other cellular protein. They used CRISPR...
  7. Hot Plasmids February 2024

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    Blog Post
    ...which uses a much shorter crRNA. Cas12-RNP-PAGE achieved high editing frequency (nearly 100% in some cases...method is fast! These editing frequencies can be achieved with a 30-minute incubation, even with multiplexed...successful in primates. The authors used a multispecies approach to generate and screen a library of AAV9 variants...best-performing variant, CAP-Mac, was effective at breaching the blood-brain barrier and delivering a transgene...
  8. CRISPR in the Clinic

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    Blog Post
    ...These therapies are delivered with an autologous approach, editing patient cells ex vivo and then returning...but especially concerning when using a systemic approach. Regarding effectiveness, if the editing frequency...meaningful way. Delivery, especially for systemic approaches, has its own unique set of obstacles. Thus far... liver. Similarly, for cardiovascular disease approaches targeting cholesterol, these particles were an...
  9. Fluorescent Proteins 101: Introduction to FRET

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    Blog Post
    ...whether two proteins are within 10 nanometers of each other, or measure the tension in the helical structure...the fluorophores must be located within 1-10nm of each other and be oriented appropriately for energy transfer...Intermolecular FRET can be experimentally difficult to achieve, because the ratio of acceptor to donor fusions...PMC2396198. 4. Sustarsic, Marko, and Kapanidis, Achillefs N. "Taking the ruler to the jungle: Single-molecule...
  10. Rabies and Neuronal Tracing

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    Blog Post
    ...way, but rather just labels the whole brain. To achieve labeling of only first-order connections to the...cell type specificity of the starter cell can be achieved by using EnvA-pseudotyped RABV, and only expressing...supplied in trans. Monosynaptic RABV spread can be achieved by modifying the wild-type RABV genome, which ...previously thought. The differences in the inputs to each of the two populations could be used to explain ...
  11. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

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    Blog Post
    ...non-functional fragments. When these two fragments are near each other, they combine forces to form a functional ...complementation assay (PCA). In the Split-BioID method, each BirA* fragment is then fused to one of two bait ...and Z interact, the two BirA* fragments complement each other to form a functional BirA* enzyme. When this.... (2017). Split-BioID a conditional proteomics approach to monitor the composition of spatiotemporally...
  12. Plasmids 101: Terminators and PolyA signals

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    Blog Post
    ... newly synthesized RNA from the transcription machinery. Terminators are found downstream of the gene ...three RNA polymerases (Polymerases I, II, and III), each responsible for transcribing different types of ...termination are understood and it has been noted that each eukaryotic RNA polymerase terminates differently.... PubMed Central PMCID: PMC104046. Wodrich H, Schambach A, Kräusslich H-G. Multiple copies of the Mason–Pfizer...
  13. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

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    Blog Post
    ... to linking pieces of DNA that are far away from each other in terms of base pairs, but close together...method is that a new cell line has to be created for each genomic loci of interest. This is cumbersome and...analyzed with CAPTURE 2.0, with two gRNAs targeting each site. They found that DNA sequences targeted by ...to interact with, and how many promoters or genes each enhancer is interacting with. Super enhancers are...
  14. The AAVantages of AAV in CRISPR Screens

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    Blog Post
    ...vectors can be made at high titers equal to those achieved with lentivirus but offer an improved safety profile...increases the possibilities when designing screens. Each serotype has its own tropism, or a preference to... you can use smaller volumes of your library for each experiment. Additional considerations One of the...Performing In Vivo CRISPR Screens Using the FITS Approach Additional resources at addgene.org CRISPR pooled...
  15. Viral Vectors 101: Voltage Indicators

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    Blog Post
    ...this later). Chemigenetic sensors take a hybrid approach and utilize both a genetic component as well as...sensing to signal output slightly slower than other approaches. Functionally, the voltage sensitivity of the...level in vitro. In vivo monitoring has also been achieved with these sensors, but further optimization is...cell level as they are in vitro. Factors like photobleaching, lack of red-shifted indicators, and brightness...
  16. Tips to Make the Most of a Scientific Conference

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    Blog Post
    ...to, I made it a personal goal to ask one question each day (this may be more or less difficult depending...identifies a potential collaborator from the meeting, reach out to them, pointing out that you heard their talk...handles to help you find other scientists attending. Reach out if you’d like to meet someone and set aside ...to set goals for myself like meet ten new people each day. Exchange business cards. When you’re done networking...
  17. CrispyCrunch: High-throughput Design and Analysis of CRISPR+HDR Experiments

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    Blog Post
    ... the human genome with fluorescent proteins. For each gene, we needed to design guide RNAs, donor templates...gaussian normalized cut-to-insert distance) This gives each guide a score ranging between 0 and 1. The cutting...target. The CFD score ranges between 0 and 100 for each guide, with 100 being the strongest interaction ...but the protein sequence they encode is not. After each mutation, it will check the CFD score. If it is ...
  18. Truncated gRNAs for Regulating Gene Expression

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    Blog Post
    ...the genome. Native Cas9 has two catalytic domains, each of which cleaves one strand of DNA upon binding ...proteins derived from different bacterial species, each of which interacts with a distinct gRNA that allows...expression with truncated gRNAs An alternative approach is to modulate Cas9’s nuclease activity by modifying...engineering parameters via a library-on-library approach." Nature methods 12.9 (2015): 823-826. PubMed ...
  19. Transferable Skills Guide: Managing a Team

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    Blog Post
    ... when I first became a manager. Know your people Each person is unique and should be managed accordingly...writing a grant, thinking about the next project, reaching out to other teams, expanding your network…. I...problem, I was able to resolve this issue - with coaching - before it became a major problem. But I also...always the case or you may not feel comfortable reaching out to your manager. This is when having a mentor...
  20. Academic vs. Industry Postdocs

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    Blog Post
    ...Genentech, will not. The point is that each company has a unique approach, and some might suit your needs better...setting—especially if job applications require teaching experience or high numbers of publications, which...into the general careers section of their web site. Each position is for a specific, pre-approved project...
Showing: 421 - 440 of 958 results